Analysis of Volatile Constituent by Hydrodistillation and Solid-Phase Microextraction Techniques and Antimicrobial and Scolicidal Activities of Essential Oil and Soxhlet Extracts of Ulva rigida grown in Turkey.

In the present study, the volatile composition of Ulva rigida (U. rigida) was elucidated by two different methods. As a result of the identification process of volatile components using the GC/MS-FID instrument, 31 compounds were identified by hydrodistillation (HD) method, and 15 compounds were identified by solid-phase microextraction (SPME) method, elucidating the structure of 99.86 % and 92.65 %, respectively. The most abundant compounds in the essential oil of U. rigida were n-hexadecanoic acid and pentadecanal, while the most abundant compound according to the SPME analysis was heptadecyne, a hydrocarbon compound. In the next step, hexane, dichloromethane, chloroform and methanol solvent extracts of U. rigida were prepared and the antimicrobial activities of the extracts and the essential oil obtained by hydro-distillation as well as the scolicidal activities of the solvent extracts were determined. The results of the antimicrobial activity test of the essential oil showed a high level of activity against Bacillus cereus ATCC 10876 and MRSA. The highest activity was found on the microorganism of Pseudomonas aeruginosa ATCC 9027 in chloroform and methanol extracts of U. rigida. Furthermore, viability detection was performed and the scolicidal effects of the extracts on protoscoleces were assessed. The values of lethal concentration doses (LD50 , LD75 and LD90 ) were calculated using probit analysis.

Analysis of mitochondrial DNA cytochrome-b (CYB) and ATPase-6 gene mutations in COVID-19 patients.

Coronavirus disease of 2019 (COVID-19) is a pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Mutations of mitochondrial DNA (mtDNA) are becoming increasingly common in various diseases. This study aims to investigate mutations in the cytochrome-b (CYB) and adenosine triphosphatase-6 (ATPase-6) genes of mtDNA in COVID-19 patients. The association between mtDNA mutations and clinical outcomes is investigated here. In the present study, mutations of the mtDNA genes CYB and ATPase-6 were investigated in COVID-19 (+) (n = 65) and COVID-19 (-) patients (n = 65). First, we isolated DNA from the blood samples. After the PCR analyses, the mutations were defined using Sanger DNA sequencing. The age, creatinine, ferritin, and CRP levels of the COVID 19 (+) patients were higher than those of the COVID-19 (-) patients (p = 0.0036, p = 0.0383, p = 0.0305, p < 0.0001, respectively). We also found 16 different mutations in the CYB gene and 14 different mutations in the ATPase-6 gene. The incidences of CYB gene mutations A15326G, T15454C, and C15452A were higher in COVID-19 (+) patients than COVID-19 (-) patients; p < 0.0001: OR (95% CI): 4.966 (2.215-10.89), p = 0.0226, and p = 0.0226, respectively. In contrast, the incidences of A8860G and G9055A ATPase-6 gene mutations were higher in COVID-19 (+) patients than COVID-19 (-) patients; p < 0.0001: OR (95%CI): 5.333 (2.359-12.16) and p = 0.0121 respectively. Yet, no significant relationship was found between mtDNA mutations and patients' age and biochemical parameters (p > 0.05). The results showed that the frequency of mtDNA mutations in COVID-19 patients is quite high and it is important to investigate the association of these mutations with other genetic mechanisms in larger patient populations.

Survey and first report of Acanthamoeba T4 genotype in natural spring water resources in the Black Sea, Turkey.

Infection with Acanthamoeba spp. may result in granulomatous amoebic encephalitis and Acanthamoeba keratitis. Water is an important habitat where Acanthamoeba species thrive. Therefore, studying the occurrence of this free-living amoeba in water sources will help understand the infection dynamics. The aim of the study was to survey and report on the presence of Acanthamoeba spp. in water resources from the Ordu and Giresun provinces in Black Sea. Acanthamoeba spp. was found in 1/17 natural spring water samples from Ordu and in 2/18 from Giresun. Acanthamoeba species were not detected in any of the investigated tap water samples. Sequencing of the (SSU) rDNA gene resulted in the identification of haplotype I (Acanthamoeba genotype: KJ094684). T4 (8.6%) was the only isolated genotype in both Ordu and Giresun provinces. This is the first report of Acanthamoeba T4 genotype in natural spring water resources in the Black Sea. The occurrence of Acanthamoeba species in natural spring water sources should be considered as a potential risk for human infection, especially to high-risk populations.

Acanthamoeba spp. in river water samples from the Black Sea region, Turkey.

The present study aims to investigate the occurrence of free living amoeba (FLA) in water resources (rivers and tap water) in Samsun in the Black Sea. The presence of Acanthamoeba spp. was confirmed in 98 of 192 water samples collected from 32 sites of Samsun province (Samsun centre, Terme, Carsamba, Tekkekoy, Bafra) by PCR. Acanthamoeba spp. were found in 15/36 river samples from Samsun, in 58/90 from Terme, in 12/30 from Carsamba, in 7/18 from Tekkekoy and in 6/18 from Bafra. No Acanthamoeba species were detected in tap water samples. The highest rate in river waters contaminated with Acanthamoeba species was in Terme followed by Samsun centre (41.7%), Carsamba (40%), Tekkekoy (38.9%) and Bafra districts (33.3%), respectively. The result of the subsequent sequence analysis showed Haplotype I (A. triangularis) in 5%, Haplotype II (A. polyphaga) in 29.6%, Haplotype III (Acanthamoeba spp.) in 62% and Haplotype IV (A. lenticulata) in 3%. The most common genotype was Acanthamoeba T4 (Acanthamoeba spp., A. polyphaga, A. triangularis) and T5 genotype was also found in 3%. The T4 genotype is the most common genotype associated with Acanthamoeba keratitis (AK) worldwide; therefore, humans and animals living in the area are at risk after contact with such waters.

[A new strategy for enhancing acanthamoebicidal activity with synthesis of nanoflower of Laurocerausus officinalis Roemer (cherry laurel) fruit extracts]. / Laurocerausus officinalis Roemer (taflan) meyve ekstrelerinden nanoçiçek sentezi ile akantamoebisidal aktivitenin arttirilmasinda yeni bir strateji.

Pathogenic Acanthamoeba species often cause infection known as Acanthamoeba keratitis among people who use contact lenses. It is a type of infection that can result in corneal ulceration, visual loss or even blindness, if not treated. There are various therapeutic options available in the treatment of Acanthamoeba infections but they are usually tough treatments with limited efficacy. For instance, hydrogen peroxide (H2O2) is a commonly used contact lens disinfectant which is effective against Acanthamoeba but it is toxic to the cornea. For these reasons, new and more efficacious treatment options are required for Acanthamoeba infections. In this context, plants are considered natural resources for the discovery of new drugs. Laurocerasus officinalis Roem. (cherry laurel) (Rosaceae) grows in Black Sea region; and it is known as "Taflan", "Laz kirazi" or "Karayemis". Local people are using the seeds against diabetes, while the fruits are consuming as food, and used fordiuretic and passing kidney stones. It has also been reported that the seeds of the cherry laurel are used as an antiparasitic agent in this area. The aim of the study was to confirm the traditionally use of antiparasitic activity of this fruit and to increase the potential effect by means of organic-inorganic hybrid synthesis. Total phenol contents of methanol extracts prepared from endocarp, mesocarp and seeds of the fruit were calculated. The effects of methanol extracts and nano flower (NFs) plants synthesized from these extracts on the proliferation of Acanthamoeba castellanii were investigated. Thus, for the first time, novel organic-inorganic nanobio-antiparasitic agents called NFs were produced from cherry laurel and the increase in the amoebicidal activity of the NFs was elucidated. The characterization of NFs were determined with Scanning Electron Microscopy (SEM), Fourier Transform Infrared Spectrometer (FT-IR) and Energy-Dispersive X-ray (EDX) techniques. In addition, the catalytic activity of the fruit extracts and the NFs were measured against guaiacol in the presence of H2O2. The viability testing of A.castellanii cysts used for amoebicidal activity was performed using 4% trypan blue. Methanol extracts and nano-flowers were prepared at concentrations of 32, 16, 8, 4, 2 and 1 mg/ml in 0.9% saline and distributed 200 µl each in tubes and incubated in the room temperature with the addition of 200 µl of 98% viable A.castellani parasites. The results were evaluated using the SPSS V.22.0 program and it was determined that there was a significant increase in the amoebicidal activity of NFs compared with the other extracts according to variance analysis (p≤ 0.05). In the study, it was determined that samples killed parasites or reduced parasite proliferation at certain times. As a result, NFs synthesized from fruit extracts were demonstrated about three times more effective than the non hybrid extracts for amoebicidal activity. This situation can be explained as high proliferative effect of a new nano-bio-antiparasitic agent known as nanoflower against A.castellanii.

PREVALENCE OF DEMODEX ECTOPARASITES AMONG HUMANS IN ORDU PROVINCE IN TURKEY.

Demodex folliculorum and Demodex brevis are common ectoparasites in humans. Demodex parasite infestations have not been determined in the province of Ordu. We determined the prevalence of Demodex species among humans in Ordu Provience, Turkey. Seven hundred ninety-nine subjects (438 males and 361 females) aged ≥ 18 years living in the central districts of Ordu Province, Turkey, were selected using the World Health Organization cluster sampling method. A superficial skin biopsy of the face was obtained from each subject. Six hundred sixty-nine subjects (83.7%) had a Demodex parasite. Factors significantly associated with the presence of Demodex infestation were: female gender, employment in the private sector, people who only occasionally wash their face and district of residence. Since Demodex ectoparasites were common in Ordu Province, it is suggested that the diagnosis and treatment of this ectoparasite should be carried out in the hospitals of this region.

[Investigation on Toxoplasma gondii by polymerase chain reaction and loop-mediated isothermal amplification in water samples from Giresun, Turkey]. / Giresun'da su örneklerinde Toxoplasma gondii'nin polimeraz zincir reaksiyonu ve ilmige dayali izotermal amplifikasyon yöntemleriyle arastirilmasi

Toxoplasmosis is generally asymptomatic in immunocompetent subjects, however serious manifestations of the disease may develop in immunocompromised patients and in pregnant women. The mean Toxoplasma gondii seroprevalence which is approximately 40% in Turkish population, indicates the high risk for the development of acute toxoplasmosis in those cases. One of the transmission ways of T.gondii is the consumption of contaminated water. The aim of this study was to detect the presence of T.gondii in the environmental and drinking water samples by using standard polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) methods. A total of 96 water samples, of them 76 were environmental water samples collected from creeks in Giresun province center and its various districts (Piraziz, Bulancak, Kesap, Espiye) and 20 from drinking water samples, were included in the study. The samples were precipitated by the aluminium sulfate method and DNAs were isolated from the pellets formed with the sucrose gradient method. PCR and LAMP were applied to the isolated DNAs, by the use of primers F3 and B3 specific to B1 gene of the parasite. In our study, T.gondii was detected in none of the drinking water samples by PCR and LAMP, however T.gondii DNAs were positive in 13.2% (10/76) of the environmental water samples. Both of the methods yielded the same results in those samples. The stations that were positive for T.gondii were Aksu creek in Giresun province center; Gelivera creek in Espiye; Yolagzi, Kesap, Kesap Login Bridge creeks in Kesap; Bulancak, Karadere and Incivez creeks in Bulancak; Piraziz and Çayiragzi creeks in Piraziz counties. Resistance of T.gondii to chlorination and the inadequacy of the filtration processes, create a serious threat among people in contact with rivers, sea and drinking waters particularly in areas with high humidity. As far as the national literature was considered, no report about the water-borne T.gondii infections were detected in Giresun province of Black Sea region, Turkey. Thus this study will aid to the literature related to water-borne T.gondii infections. The results of this study emphasizes the essential need for appropriate disinfection procedures and purification processes before the discharge of waste water to prevent the cases of water-borne toxoplasmosis.

[Tuboovarian abscess caused by hydatid cyst: a rare case]. / Nadir bir olgu: Tuboovaryan apseye neden olan hidatik kist.

Primary lesions of hydatid cysts caused by Echinococcus granulosus, are frequently localized in liver, followed by lungs, muscles, kidneys, spleen and bones. Pelvic inoculations are rare and usually occur as a secondary infection. In this report, a case of primary hydatid cyst in the abdomen, spleen and pelvic organs, clinically mimicking tuboovarian abscess, was presented. A nineteen-years-old female patient was admitted to the gynecology outpatient clinic with the complaint of abdominal pain for two days. The case was considered as tuboovarian abscess according to the initial examination findings and hospitalized for treatment and follow-up. In transabdominal ultrasound examination, 44 x 43 mm thin-walled septated cysts in the left ovary and 65 x 65 mm thin-walled multiloculated cysts in the spleen were detected. Abdominal computerized tomography also yielded multivesicular cystic masses in spleen, front abdominal wall and the left ovary. Since the clinical and vital findings worsened, she initially underwent ovarian cystectomy by laparoscopy, then abdominal cystectomy and splenectomy. The operation material examined macroscopically was compatible with hydatid cyst with the characteristics of a germinative membrane and hydatid sand. The diagnosis was confirmed by histopathological examination. The patient was discharged without complication on post-operative sixth day, with a recommendation of albendezol (15 mg/kg/day, 3 months) treatment. Since the patient had undergone emergency surgery, indirect hemaglutination (IHA) test had not been performed pre-operatively. However, post-operative third month serum sample revealed a positive (1/32) IHA titer. In conclusion, hydatid cyst should be kept in mind in the differential diagnosis of patients with abdominal pain, in response to the high prevalence of the parasite in our country.

Do antidiabetic drugs prevent the transformation of Acanthamoeba trophozoite into cyst form?

This study examines the effects of three different drugs with metformin, acarbose and pioglitazone active ingredients used for antidiabetic purposes on Acanthamoeba cysts and trophozoites. Cultures of A. castellanii trophozoites and cysts were prepared to test the anti-amoebic activity of metformin, acarbose and pioglitazone. Cultures were then prepared for A. castellanii cyst and trophozoite forms and parasites were exposed to different concentrations (0.750 mg/mL, 0.375 mg/mL, 0.186 mg/mL and 0.093 mg/mL) of metformin, acarbose and pioglitazone. As a result of the study, the reproductive potential suppressive effects and conversion from trophozoite form to cyst form of all three substances on A. castellanii trophozoites and cysts were determined. Parasites were counted at 12, 24 and 48 hours in the cell counter after staining with trypan blue. In comparison of the effects of metformin, acarbose and pioglitazone used in the study on A. castellanii trophozoites and cysts, it was observed that all three substances were statistically effective against cysts and trophozoites at a concentration of 0.750 mg/mL. Furthermore, it was determined that all concentrations of the three active substances included in the study significantly decreased the rate of cyst formation even at the end of the 7th day. In this context, it was determined that all three substances have amebicidal effects, and they significantly inhibit the transformation of A. castellanii trophozoites to cyst form. It is thought that these active substances, which are currently used as anti-diabetic, can be used in combination with other drugs in A. castellanii infections based on our study findings.

Prevalence of House Dust Mites and Presence of Der p 1 and Der f 1 in Ordu, Giresun, Trabzon and Rize Provinces.

OBJECTIVE: This study was carried out to detect house dust mites in houses and to investigate group 1 antigens of Dermatophagoid species in Ordu, Giresun, Trabzon and Rize provinces of the Central and Eastern Black Sea Region. METHODS: Dust samples obtained from the beds were subjected to both microscopic and antigenic examination. Samples prepared by the lactic acid method for microscopic examination were evaluated under a light microscope. Antigenic analysis was performed by investigating Der p 1 and Der f 1 belonging to D. pteronyssinus and D. farinae by ELISA test. RESULTS: 90.3% of the dust samples were evaluated positive by microscopic examination (10x, 40x) and 149 mites were detected. D. pteronyssinus 74%, D. farinae 13%, Dermatophagoides spp. growth forms 5%, Cheyletus spp. 1%, E. maynei 1%, C. arcuatus 1%, T. putrescentiae 1%, L. destructor 1% and unidentified mites were detected at the rate of 3% respectively. Der p 1 antigen was detected in 93% and Der f 1 antigen in 84.7%. The highest amount of antigen detected in one gram of powder was 1,272 µg for Der p 1 and 0,482 µg for Der f 1. CONCLUSION: No difference was observed between mite species and distribution in the provinces where the study was conducted (p<0.05). Dermatophagoides were found in 93% of the population. The low (4%) rate of storage/food mites is related to the fact that samples were not taken from the floors. Antigen accumulation may be important in the beds since the activity of the mites is observed throughout the year in temperate and humid regions. It is thought that this diagnosis method can be used and can be taken into account in terms of the environments in which sensitive people live.

Investigation of Demodex Spp. prevalence among managers and workers of health hazard bearing and sanitary establishment.

BACKGROUND/PURPOSE: Two Demodex species are known to live on people. Demodex folliculorum lives in the openings of hair follicles alone or in groups. D brevis lives in the depths of the sebaceous glands alone. There are different related on the epidemiology of Demodex species have been published. METHOD: In this study, taking into account that the parasite is transmitted through close contact from person to person, we aimed to evaluate the relation between the presence of Demodex spp with gender and age among the Health hazard bearing and sanitary establishment operators and workers who came for porter examination. RESULTS: For this purpose, 862 male and 215 female patients with a total of 1077 volunteers from the face area of the standard superficial skin biopsy (SSSB) method and studied samples were taken. More than five mites in 1 cm(2) density was defined as positive. In investigated samples 37.3% Demodex spp. positivity was detected. Only two were found to be D brevis and the others were D folliculorum. The study revealed statistically significant relationships between the positivity of parasites with the occupational group, age and sex. CONCLUSION: Since the prevalence of Demodex among healthy individuals without any complaint was found to be 37%, we believe Demodex spp should be investigated in porter examinations of people who have dermatological complaints.

[Investigation of Microsporidia prevalence by different staining methods in cases of diarrhea]. / Ishalli Olgularda Microsporidia Sikliginin Farkli Boyama Yöntemleriyle Arastirilmasi

Microsporidia, depending on their different species, generally lead to self-limited, sporadic and mild infections such as diarrhea, corneal ulcer and myositis. They are considered as opportunistic pathogens in HIV-positive patients however in recent years Microsporidia have been detected also in immunocompetent individuals as a cause of diarrhea. Diagnosis of Microsporidia depends on the detection of spores or different developmental stages of protozoon in stool, urine, sinus aspirates, nasal discharge, bronchoalveolar lavage or tissue biopsies. Diagnosis of Microsporidia infections is usually achieved by the use of different staining methods, serological tests, polymerase chain reaction, and electron microscopic methods. The aims of this study were to detect the incidence of microsporidia in patients with diarrhea by using three different staining methods and to compare the performance of these methods. A total of 225 stool samples from diarrheal patients (84 were children, 141 were adults; 103 were female, 122 were male) admitted to Gazi University Medical Faculty Hospital between March-June 2009, have been evaluated in the laboratory of Medical Microbiology Department. Stool samples were examined in terms of the presence of Microsporidia spores by Weber's modified trichrom staining (MTS), calcofluor (CF) and acridine orange (AO) staining methods. Microsporidia positivity rate was 9.8% (22/225) in the diarrheal patients, the rate being 9.5% (8/84) in children and 9.9% (14/141) in adults. There was no statistically significant difference between age and gender groups (p> 0.05) regarding Microsporidia detection. When MTS was considered as the reference method, sensitivity, specifity and consistency of AO staining were estimated as 100%, 91.6% and 92%, respectively, while those rates for CF staining were 95.4%, 99.5% and 99%, respectively. There was very strong and significant correlation (r= 0.950, p< 0.001) between CF staining and MTS, while there was strong and significant (r= 0.719, p< 0.001) correlation between AO staining and MTS. Although AO staining is rapid and convenient, the positive predictive value was measured very low (56.4%) and the interpretation of stained slides was very difficult since background of the slides was stained orange and there were a lot of dye artefacts. In conclusion, screening Microsporidia in all diarrheal stool samples is of diagnostic value. To increase sensitivity and reliability in the detection of Microsporidia spores in diarrheal samples, initial application of calcofluor staining should be followed by the confirmatory MTS method.

[The rates of seropositivity and seroconversion of toxoplasma infection in pregnant women]. / Gebelerde Toksoplazma Enfeksiyonunun Seropozitiflik ve Serokonversiyon Oranlari

Infections caused by Toxoplasma gondii are frequently asymptomatic in healthy adults, however they may be serious in pregnant women and immunocompromised patients. The aims of this study were to investigate the rates of seropositivity and seroconversion in pregnant women and newborn cord blood samples, and to evaluate those data in the view of relation to lifestyle and nutrition. A total of 312 pregnant women (mean age: 28.1 ± 5.2 years) who were admitted to and followed by gynecology clinics of Inonu University Medical School Hospital, Malatya, Turkey were included in this observational and cross-sectional study. Anti-toxoplasma IgG and IgM antibodies in pregnants and newborn cord sera were screened by commercial ELISA and immunofluorescence antibody (BioTek; USA) methods. A total of 312 sera from pregnant women and 312 cord blood samples during delivery were collected. IgG seropositivity rate in pregnants was found as 37.5% (117/312), seroconversion was not determined in restrained pregnants and T.gondii IgM was found negative in all pregnants. Also in all newborns IgM was negative and IgG seropositivity was determined as 33.3% (104/312) in cord blood. There was a statistically significant relationship between IgG seropositivity and raw meat consumption (p< 0.001) and being engaged in agriculture (p< 0.005). It was concluded that toxoplasma antibodies should routinely be searched on the first visit of the pregnants and the seronegative cases should be trained about the preventive measures related to toxoplasmosis. The follow-up of toxoplasma seronegative cases during pregnancy can be achieved by only detecting the IgM class antibodies and this will also reduce the cost of screen test.

Comparative Evaluation of Re 529-Bp Sequence and B1 Gene in the Detection of Toxoplasma gondii Through PCR in Water Samples of Denizli, Turkey.

PURPOSE: While Toxoplasma gondii (T. gondii) infection is asymptomatic in immunocompetent individuals, it is a life-threatening protozoan in immunocompromised individuals. Its water-borne transmission to humans poses a serious public health concern. Polymerase Chain Reaction (PCR) has a considerable potential for the sensitive and specific detection of T. gondii oocysts in waters. METHODS: Comparative evaluation of RE 529-bp sequence and B1 gene to detect T. gondii tachyzoites and oocysts via PCR in agricultural irrigation water taken from downtown Denizli, Turkey and water samples collected from neighborhood fountains was performed for the first time in Turkish context. RESULTS: Based on real-time PCR targeting the B1 genetic markers and RE 529-bp sequence, T. gondii DNA was identified in 6 (16.7%) out of 48 samples collected from agricultural irrigation water. Besides, our PCR analysis did not establish any presence of T. gondii in drinking water samples. CONCLUSION: T. gondii showed lower sensitivity in B1-based PCR than in PCR targeting RE 529-bp sequence.

The Presence of Protozoan Parasites in Drinking Waters and Environmental Waters in Denizli City Center

Objective: The presence of protozoan parasites in agricultural irrigation and drinking water resources at Denizli city center was investigated in detail for the first time. Methods: The research was carried out between October 2017 and October 2018 and 84 water samples were taken from 7 different stations identified from the Denizli city center. After examining the samples by direct visualization (Native-Lugol), they were stained with quinton's acid fast, giemsa and trichrome dyes. The preparations were evaluated parasitologically under a light microscope. Results: Cryptosporidium spp. was detected in 21 (25%) of 36 agricultural irrigation water samples collected during the study, Cyclospora cayetanensis in 5 samples (5.95%) and Giardia spp. in 12 samples (14.28%). No parasite findings were found in any of the 48 drinking water samples collected. Conclusion: The widespread use of animal husbandry and agriculture as grazing land in the sampling stations, the mixing of domestic wastewater into these waters without any treatment, and seasonal conditions cause the protozoan parasites to be seen more in certain periods. It is thought that waterborne protozoan infections that may occur in the future can be significantly prevented by taking the necessary precautions in terms of public health and environmental animal husbandry.

Comparison of the multi-epitope recombinant antigen DIPOL and hydatid fluid for the diagnosis of patients with cystic echinococcosis.

The use of serological tests containing multiple immunodominant antigens rather than single antigens have the potential to improve the diagnostic performance in Cystic Echinococcoses (CE) as a complement tool to clear the inconclusive imaging data. Here, we comparatively evaluated the diagnostic value of Hydatid Fluid (HF) and the recently described recombinant multi-epitope antigen DIPOL in IgG-ELISA in a clinically defined cohort of CE patients. The serum samples from 149 CE patients were collected just before surgical or Percutaneous- Aspiration- Injection- Reaspiration (PAIR) procedures. Additionally, serum samples of patients with other parasitic infections (n=49) and healthy individuals (n=21) were also included in the study as controls. To investigate the association between the genotype of the parasite and DIPOL, cyst materials from 20 CE patients were sequenced. In terms of overall sensitivity, HF was higher than DIPOL (82.55%,78.52%, respectively). However, while the sensitivity of HF was higher than DIPOL in patients with active and transitional cysts (83.3%, 75.4%, respectively), sensitivity of DIPOL in inactive cysts was higher compared to HF (95.6%, 78.3%, respectively). The sensitivity of DIPOL depending on cyst stage was statistically significant (P= 0.041). In terms of specificity, DIPOL was found to be better than HF (97.71%, 91.43%, respectively). By genotyping, the majority of 20 patients showed G1 genotype (80%). All patients harboring G3 and G1/G3 cyst genotypes were positive with both antigens, while 87.5% of patients with G1 genotype were seropositive with HF and 75% with DIPOL. The overall sensitivity and high specificity of DIPOL suggest that this recombinant protein containing immunodominant epitopes is a potential substitute for the HF by serological tests for the diagnosis of CE.

[Investigation of microsporidia in patients with acute and chronic urticaria]. / Akut ve Kronik Ürtikerli Hastalarda Microsporidia Pozitiflik Orani

Microsporidia species are one of the major causes of severe diarrhea especially in immunocompromised patients, however they also cause infections in immunocompetent subjects. Urticaria, which is a common dermatologic disease may be triggered by drugs, infections, foods or food additives, psychogenic factors and autoimmune, metabolic and malignant diseases. While the etiologic grounds of acute urticaria are generally identified, the etiology remains unknown in most of the chronic urticaria cases. The studies on the roles of parasitic infections in the etiology of urticaria have indicated that the most responsible protozoa are Giardia intestinalis and Blastocystis hominis. However, no data have been found in the literature concerning the relationship between Microsporidia and urticaria. The aim of this study was to investigate the presence of Microsporidia spores in the stool samples of patients diagnosed as acute or chronic urticaria in dermatology clinics and to determine the rates of positivity. All of the samples stained with modified trichrome and calcofluor staining methods were examined microscopically. The samples were accepted as positive when the spores of Microsporidia were detected by both of the staining methods. Microsporidia were detected in 26 (19.7%) of the patients and in 1 (2.8%) of the control subjects. Thus the total rate of positivity was 16% (27/168). In cases with acute/chronic urticaria, microsporidia positivity rate was found significantly higher than the control group (p= 0.028; p< 0.05). No statistically significant difference was detected between Microsporidia positivity and age or gender (p= 0.27 and p= 0.99, respectively; p> 0.05). In conclusion, Microsporidia should be taken into consideration in patients with unknown origin of urticaria. However, advanced studies are needed for supporting the relation between Microsporidia and acute/chronic urticaria.

Single-strand conformation polymorphism-based genetic characterization of the Cyclospora cayetanensis strains collected from different provinces in Turkey.

INTRODUCTION AND OBJECTIVE: Cyclospora cayetanensis, a coccidian protozoan species, has been recently found to cause diarrhea in all age groups in immunocompetent and immunocompromised individuals in most regions of the world. This study aimed to conduct the molecular detection of C. cayetanensis and to determine the genetic diversity of the 18S ribosomal RNA (rRNA) gene sequence of C. cayetanensis isolated from individuals living in different provinces in Turkey by using PCR-single-strand conformation polymorphism (SSCP). MATERIAL AND METHODS: A total of 22 subjects were included in the study. Fourteen of the subjects were female and eight were male, with ages ranging between 7-65 years. Stool specimens were examined using wet mount and modified acid-fast staining methods, which revealed the presence of oocysts in the samples. The 18S rRNA ITS-1 Ccits37f-GCTTGCTATGTTTTAGCATGTGG and Ccits501r-GCACAATGAATGCACACACA gene regions were used as primers. The PCR products were analyzed by agarose gel electrophoresis and visualized on a UV transilluminator. For the SSCP, the PCR products were denatured with formamide, run for 16 h in 6% (49:1) polyacrylamide gel, and then imaged with silver staining. RESULTS: SSCP assay was performed given that the DNA strands demonstrated different folds; the DNA strands contain different nucleotides based on the PCR-SSCP results for the Cyclospora strains collected in 4 provinces. Moreover, 3 different band profiles were observed in the investigated samples. A slight mutation difference was observed among the strains collected. CONCLUSIONS: Further comprehensive studies involving more C. cayetanensis-positive specimens and utilizing different mutation screening methods are warranted to demonstrate mutation differences in Cyclopora strains in Turkey.

[Malondialdehyde, glutathione and nitric oxide levels in patients with Enterobius vermicularis infection]. / Enterobius vermicularis ile enfekte hastalarda malondialdehid, glutatyon ve nitrik oksit düzeyleri.

The aim of this study was to investigate the levels of malondialdehyde (MDA) and nitric oxide (NO) which are indicators of oxidative stress and also the level of glutathione (GSH) which is an antioxidant molecule, in patients with Enterobius vermicularis infection. A total of 41 patients (mean age: 36.4 years; 31 were male) and 40 healthy controls (mean age: 40.2 years; 28 were male) were included to the study. None of the patients and controls have had history of hormone/steroid drug use, smoking and alcoholism. The mean level of GSH in patient and control groups were found 1.17 +/- 0.02 micromol/l and 2.49 +/- 0.10 micromol/l; MDA 26.97 +/- 2.06 micromol/l and 19.47 +/- 2.25 micromol/l; NO 20.74 +/- 0.60 micromol/dl and 17.83 +/- 0.50 micromol/dl, respectively. The mean GSH level in patient group was statistically significantly lower (p < 0.05) than controls, while the mean MDA and NO levels were statistically significantly higher (p < 0.05). These results indicated that the consumption of GSH was increased due to the severity of oxidative stress in patients infected with E. vermicularis. Detailed experimental and clinical studies are required to enlighten the relation of GSH in the pathogenesis of E. vermicularis infection. Since oxidative stress is increased during enterobiosis, the use of antioxidant agents (e.g. vitamins C and E) for the supportive treatment deserves evaluation.

Role of Adenosine Deaminase in Patients with Erythematotelangiectatic Rosacea and Demodex folliculorum Positivity.

BACKGROUND: Adenosine deaminase (ADA) is an aminohydrolase involved in the catabolism of purine nucleotides and irreversibly deaminizes adenosine and deoxyadenosine to inosine and deoxyinosine. ADA enzyme deficiency results in the loss of functional properties of B and T lymphocytes. Demodex species have been reported to be transmitted between humans through close contact and to play a role in the pathogenesis of rosacea, acne vulgaris, perioral dermatitis, seborrhoeic dermatitis, micropapillary-pruritic dermatitis and blepharitis. The present study aimed to compare serum ADA levels in D. folliculorum positive patients with the healthy control individuals. METHODS: Serum ADA levels were examined for 30 patients diagnosed with erythematotelangiectatic rosacea and 40 healthy individuals in Malatya Inonu University in 2017. Standardized skin surface biopsy (SSSB) method was used to diagnose D. folliculorum. A significant decrease was found in the ADA levels of Demodex-positive rosacea patients when compared to the control group. RESULTS: ADA levels were decreased in the Demodex-positive group. The mean ADA level in patient group was significantly lower than the mean in the control group (P<0.001). There was no significant difference between the patient and control groups in terms of age and gender. CONCLUSION: During and after treatment of Demodex-positive rosacea patients, determination of ADA levels may give more detailed information on the immune mechanisms.